心肌缺血预适应及后适应保护中Hes1配体蛋白的差异蛋白组学研究

大量研究已证实缺血预适应/后适应(IPC/I-postC)的内源性心肌保护作用，为缺血再灌注损伤的基础研究指明了新的方向，同时也为缺血性心肌病的临床治疗提供了新的契机。在总结前人与自身工作基础上，我们假设：IPC/I-postC保护作用可能通过Hes1及其配体蛋白调控相关的信号通路，并可能影响这些通路的下游，完全可能作为其共同环节。本项目拟将串联亲和纯化/二维蛋白电泳/基质辅助激光解吸及电离飞行质谱分析法等蛋白质组学技术有机结合，筛选典型的差异性高表达Hes1配体蛋白X。通过体外、细胞学实验确认其与Hes1的相互作用，探讨Hes1对其下游配体蛋白及其相关信号转导通路的影响，并通过蛋白组学技术进一步研究配体蛋白X的甲基化、磷酸化、乙酰化等改变对IPC/I-postC保护效应的影响。为进一步阐明IPC/I-postC保护效应的分子生物学机制奠定理论与实验基础，为缺血性心脏病的诊疗提供理论基础。

A large number of researches have confirmed the endogenous myocardial protective effects of ischemic preconditioning (IPC) and ischemic postconditioning (I-postC), which points out the new direction for the basic research of ischemia reperfusion injury and provides a new opportunity for the clinical treatment of the ischemic heart diseas. On the basis of predecessors' and our own works, we assume that: the IPC/I-postC protection may regulate the related signaling pathways through Hes1 and its partner-proteins, influence the downstream pathways of these pathways, and maybe the common link of these pathways. Our project will combine the proteomics technologies, such as the tandem affinity purification (TAP)/two-dimensional electrophoresis (2-DE)/matrix assisted laser desorption ionization-time of flight-mass spectrometer (MALDI-TOF-MS) analysis, screen a typical high expression protein X of the Hes1 partner-proteins. We will confirm its interaction with Hes1 by in vitro and cytological experiments, investigate the influence of Hes1 to its downstream partner-proteins and related signal pathways, and do the further researches on the methylation, phosphorylation, acetylation of the partner-protein X and the influence of the IPC/I-postC protective effect via proteomics technology. In order to establish the theoretical and experimental basis of the molecular biology mechanisms on IPC/I-postC protective effects, and provide the theoretical basis for diagnosis and treatment of the ischemic heart disease.